The present invention relates to high molecular weight Dermatophagoides proteins, nucleic acid molecules and therapeutic and diagnostic reagents derived from such proteins.
Immunoglobulin E (IgE) mediated allergic symptoms afflict many animals. IgE antibody production in an animal can induce pathogenic IgE responses including, for example, atopic disease, asthma and rhinitis. Allergens are proteins or peptides characterized by their ability to induce a pathogenic IgE response in susceptible individuals.
House dust mite (e.g., Dermatophagoides farinae and Dermatophagoides pteronyssinus; Der f and Der p, respectively) allergens are major causative agents associated with IgE-mediated pathogenesis. Previous investigators have identified two major groups of dust mite allergens in humans, group I (Der fI and Der p I, Mr 25,000) and group 2 (Der fII and Der p II, Mr 14,000); reviewed in Chapman, et al., Allergy, vol. 52, pp.37-379, 1997. Prior investigators have disclosed nucleotide and/or amino acid sequences for: Der fI, Der fII, Der p I and Der p II, U.S. Pat. No. 5,552,142, to Thomas et al., issued Sep. 3, 1996, U.S. Pat. No. 5,460,977, to Ando et al., issued Oct. 24, 1995, PCT Patent Publication No. WO 95/28424, by Chen et al., published Oct. 26, 1995, U.S. Pat. No. 5,433,948, to Thomas et al., issued Jul. 18, 1995, PCT Patent Publication No. WO 93/08279, by Garmen et al., published Mar. 4, 1993, or Chapman, ibid.; Derp III, PCT Patent Publication No. WO 95/15976, by Thomas et al., published Jun. 15, 1995; Der p VII, PCT Patent Publication No. WO 94/20614, by Thomas et al., published Sep. 15, 1994; a 40-kilodalton (kd) Der f allergen, U.S. Pat. No. 5,405,758, to Oka et al., issued Apr. 11, 1995, U.S. Pat. No. 5,314,991, to Oka et al., issued May 24, 1994; a 70-kd Der f allergen which is a heat shock protein (Hsp70), Aki et al., J. Biochem., vol. 115, pp. 435-440, 1994; or Noli et al., Vet. Immunol. Immunopath., vol. 52, pp. 147-157, 1996; and a 98-kd Der f paramyosin-like allergen, Tsai et al, J. Allergy Clin. Immunol., vol. 102, pp. 295-303, 1998. None of these published sequences indicates, suggests or predicts any of the mite allergic nucleic acid molecules or proteins of the present invention, nor the relevance of such proteins as being immunoreactive with IgE antibodies in canine, feline, or human sera.
Products and processes of the present invention are needed in the art that provide specific detection and treatment of mite allergy.
The present invention relates to novel proteins having molecular weights of about 60 kilodaltons (kd or kD), 70 kD, or from about 98 kD to about 109 kD. Such proteins include at least one epitope of a protein allergen of a mite of the genus Dermatophagoides and are designated herein as Der HMW-map proteins. Preferred proteins are Dermatophagoides farinae or Dermatophagoides pteronyssius proteins. The present invention also provides proteins that are fragments or peptides of full-length or mature proteins, as well as antibodies, mimetopes or muteins of any of such proteins. The present invention also provides nucleic acid molecules encoding any of such proteins, as well as complements thereof. The present invention also includes methods to obtain such proteins, nucleic acid molecules, antibodies, mimetopes or muteins, as well as methods to use such compounds in diagnostic or therapeutic applications. The present invention also relates to reagents comprising non-proteinaceous epitopes that bind to IgE in mite-allergic dogs and/or cats as well as to antibodies raised against such epitopes. The present invention also relates to therapeutic compositions or assay kits comprising such non-proteinaceous epitopes, as well as to methods to identify and/or desensitize an animal susceptible to an allergic response to a mite, comprising the use of non-proteinaceous epitopes of the present invention.
One embodiment of the present invention is at least one of the following isolated nucleic acid molecules: (a) a nucleic acid molecule comprising at least about 150 nucleotides, wherein such a nucleic acid molecule hybridizes, in a solution comprising 1xc3x97SSC and 0% formamide, at a temperature of about 50xc2x0 C., to a nucleic acid molecule comprising at least one of the following nucleic acid sequences: SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:45, and a nucleic acid sequence encoding a protein comprising the amino acid sequence of SEQ ID NO:33 and a complement thereof; and (b) a nucleic acid molecule comprising a fragment of any of the nucleic acid molecules of (a) wherein the fragment comprises at least about 15 nucleotides. The present invention also includes recombinant molecules, recombinant viruses and recombinant cells comprising such nucleic acid sequences as well as methods to produce them.
Another embodiment of the present invention is an isolated protein encoded by at least one of the following nucleic acid molecules: (a) a nucleic acid molecule comprising at least about 150 nucleotides, wherein such a nucleic acid molecule hybridizes, in a solution comprising 1xc3x97SSC and 0% formamide, at a temperature of about 50xc2x0 C., to a nucleic acid molecule comprising at least one of the following nucleic acid sequences: SEQ ID NO:16, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:36, SEQ ID NO:39, SEQ ID NO:42, SEQ ID NO:45, and a complement of a nucleic acid sequence encoding a protein comprising the amino acid sequence SEQ ID NO:33; and (b) a nucleic acid molecule comprising a fragment of any of the nucleic acid molecules of (a), wherein the fragment comprises at least about 15 nucleotides. An isolated protein of the present invention can also be encoded by a nucleic acid molecule that hybridizes under stringent hybridization conditions with the complement of a nucleic acid molecule that encodes a protein having at least one of the following amino acid sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:18, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:38, SEQ ID NO:41, and SEQ ID NO:44. The present invention also includes an antibody that selectively binds to a protein of the present invention as well as methods to produce and use such proteins or antibodies.
The present invention also includes a therapeutic composition for treating an allergic response to a mite. Such a therapeutic composition includes at least one of the following desensitizing compounds: (a) an isolated nucleic acid molecule of the present invention; (b) an isolated mite allergenic protein of the present invention; (c) a mimetope of such a mite allergenic protein; (d) a mutein of such a mite allergenic protein; (e) an antibody to such a mite allergic protein; and (f) an inhibitor of binding of such a mite allergic protein to IgE. Also included is a method to desensitize a host animal to an allergic response to a mite. Such a method includes the step of administering to the animal a therapeutic composition of the present invention.
One embodiment of the present invention is an assay kit for testing if an animal is susceptible to or has an allergic response to a mite. Such a kit includes an isolated protein of the present invention and a means for determining if the animal is susceptible to or has that allergic response. Such a means includes use of such a protein to identify animals susceptible to or having allergic responses to mites. The present invention also includes a method to identify an animal susceptible to or having an allergic response to a mite. Such a method includes the steps of: (a) contacting an isolated protein of the present invention with antibodies of an animal; and (b) determining immunocomplex formation between the protein and the antibodies, wherein formation of the immunocomplex indicates that the animal is susceptible to or has such an allergic response.
The present invention includes a reagent that comprises a non-proteinaceous epitope having at least one of the following identifying characteristics: (a) the epitope is resistant to xcex2-elimination of peptides; (b) the epitope is resistant to Proteinase-K digestion; and (c) the epitope is reactive to a test designed to detect glycosylated proteins. Such an epitope binds to at least one of the following antibodies: canine IgE from dogs allergic to mites and feline IgE from cats allergic to mites. Also included is an isolated antibody that selectively binds such a non-proteinaceous epitope as well as derivatives of such an epitope.
The present invention also relates to therapeutic compositions and assay kits comprising a non-proteinaceous epitope of the present invention, as well as methods to identify and/or desensitize an animal susceptible to an allergic response to a mite, comprising the use of a non-proteinaceous epitope of the present invention.